The Dual Activity Responsible for the Elongation and Branching of β-(1,3)-Glucan in the Fungal Cell Wall

ABSTRACT β-(1,3)-Glucan, the major fungal cell wall component, ramifies through β-(1,6)-glycosidic linkages, which facilitates its binding with other cell wall components contributing to proper cell wall assembly. Using Saccharomyces cerevisiae as a model, we developed a protocol to quantify β-(1,6)...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Vishukumar Aimanianda, Catherine Simenel, Cecile Garnaud, Cecile Clavaud, Rui Tada, Lise Barbin, Isabelle Mouyna, Christoph Heddergott, Laura Popolo, Yoshikazu Ohya, Muriel Delepierre, Jean-Paul Latge
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	Saccharomyces cerevisiae cell wall beta-glucan remodeling Aspergillus fumigatus Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/7c42e07ba98241b8ae37693ee5b262ae
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:7c42e07ba98241b8ae37693ee5b262ae
record_format	dspace
spelling	oai:doaj.org-article:7c42e07ba98241b8ae37693ee5b262ae2021-11-15T15:51:30ZThe Dual Activity Responsible for the Elongation and Branching of β-(1,3)-Glucan in the Fungal Cell Wall10.1128/mBio.00619-172150-7511https://doaj.org/article/7c42e07ba98241b8ae37693ee5b262ae2017-07-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00619-17https://doaj.org/toc/2150-7511ABSTRACT β-(1,3)-Glucan, the major fungal cell wall component, ramifies through β-(1,6)-glycosidic linkages, which facilitates its binding with other cell wall components contributing to proper cell wall assembly. Using Saccharomyces cerevisiae as a model, we developed a protocol to quantify β-(1,6)-branching on β-(1,3)-glucan. Permeabilized S. cerevisiae and radiolabeled substrate UDP-(14C)glucose allowed us to determine branching kinetics. A screening aimed at identifying deletion mutants with reduced branching among them revealed only two, the bgl2Δ and gas1Δ mutants, showing 15% and 70% reductions in the branching, respectively, compared to the wild-type strain. Interestingly, a recombinant Gas1p introduced β-(1,6)-branching on the β-(1,3)-oligomers following its β-(1,3)-elongase activity. Sequential elongation and branching activity of Gas1p occurred on linear β-(1,3)-oligomers as well as Bgl2p-catalyzed products [short β-(1,3)-oligomers linked by a linear β-(1,6)-linkage]. The double S. cerevisiae gas1Δ bgl2Δ mutant showed a drastically sick phenotype. An ScGas1p ortholog, Gel4p from Aspergillus fumigatus, also showed dual β-(1,3)-glucan elongating and branching activity. Both ScGas1p and A. fumigatus Gel4p sequences are endowed with a carbohydrate binding module (CBM), CBM43, which was required for the dual β-(1,3)-glucan elongating and branching activity. Our report unravels the β-(1,3)-glucan branching mechanism, a phenomenon occurring during construction of the cell wall which is essential for fungal life. IMPORTANCE The fungal cell wall is essential for growth, morphogenesis, protection, and survival. In spite of being essential, cell wall biogenesis, especially the core β-(1,3)-glucan ramification, is poorly understood; the ramified β-(1,3)-glucan interconnects other cell wall components. Once linear β-(1,3)-glucan is synthesized by plasma membrane-bound glucan synthase, the subsequent event is its branching event in the cell wall space. Using Saccharomyces cerevisiae as a model, we identified GH72 and GH17 family glycosyltransferases, Gas1p and Bgl2p, respectively, involved in the β-(1,3)-glucan branching. The sick phenotype of the double Scgas1Δ bgl2Δ mutant suggested that β-(1,3)-glucan branching is essential. In addition to ScGas1p, GH72 family ScGas2p and Aspergillus fumigatus Gel4p, having CBM43 in their sequences, showed dual β-(1,3)-glucan elongating and branching activity. Our report identifies the fungal cell wall β-(1,3)-glucan branching mechanism. The essentiality of β-(1,3)-glucan branching suggests that enzymes involved in the glucan branching could be exploited as antifungal targets.Vishukumar AimaniandaCatherine SimenelCecile GarnaudCecile ClavaudRui TadaLise BarbinIsabelle MouynaChristoph HeddergottLaura PopoloYoshikazu OhyaMuriel DelepierreJean-Paul LatgeAmerican Society for MicrobiologyarticleSaccharomyces cerevisiaecell wallbeta-glucanremodelingAspergillus fumigatusMicrobiologyQR1-502ENmBio, Vol 8, Iss 3 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Saccharomyces cerevisiae cell wall beta-glucan remodeling Aspergillus fumigatus Microbiology QR1-502
spellingShingle	Saccharomyces cerevisiae cell wall beta-glucan remodeling Aspergillus fumigatus Microbiology QR1-502 Vishukumar Aimanianda Catherine Simenel Cecile Garnaud Cecile Clavaud Rui Tada Lise Barbin Isabelle Mouyna Christoph Heddergott Laura Popolo Yoshikazu Ohya Muriel Delepierre Jean-Paul Latge The Dual Activity Responsible for the Elongation and Branching of β-(1,3)-Glucan in the Fungal Cell Wall
description	ABSTRACT β-(1,3)-Glucan, the major fungal cell wall component, ramifies through β-(1,6)-glycosidic linkages, which facilitates its binding with other cell wall components contributing to proper cell wall assembly. Using Saccharomyces cerevisiae as a model, we developed a protocol to quantify β-(1,6)-branching on β-(1,3)-glucan. Permeabilized S. cerevisiae and radiolabeled substrate UDP-(14C)glucose allowed us to determine branching kinetics. A screening aimed at identifying deletion mutants with reduced branching among them revealed only two, the bgl2Δ and gas1Δ mutants, showing 15% and 70% reductions in the branching, respectively, compared to the wild-type strain. Interestingly, a recombinant Gas1p introduced β-(1,6)-branching on the β-(1,3)-oligomers following its β-(1,3)-elongase activity. Sequential elongation and branching activity of Gas1p occurred on linear β-(1,3)-oligomers as well as Bgl2p-catalyzed products [short β-(1,3)-oligomers linked by a linear β-(1,6)-linkage]. The double S. cerevisiae gas1Δ bgl2Δ mutant showed a drastically sick phenotype. An ScGas1p ortholog, Gel4p from Aspergillus fumigatus, also showed dual β-(1,3)-glucan elongating and branching activity. Both ScGas1p and A. fumigatus Gel4p sequences are endowed with a carbohydrate binding module (CBM), CBM43, which was required for the dual β-(1,3)-glucan elongating and branching activity. Our report unravels the β-(1,3)-glucan branching mechanism, a phenomenon occurring during construction of the cell wall which is essential for fungal life. IMPORTANCE The fungal cell wall is essential for growth, morphogenesis, protection, and survival. In spite of being essential, cell wall biogenesis, especially the core β-(1,3)-glucan ramification, is poorly understood; the ramified β-(1,3)-glucan interconnects other cell wall components. Once linear β-(1,3)-glucan is synthesized by plasma membrane-bound glucan synthase, the subsequent event is its branching event in the cell wall space. Using Saccharomyces cerevisiae as a model, we identified GH72 and GH17 family glycosyltransferases, Gas1p and Bgl2p, respectively, involved in the β-(1,3)-glucan branching. The sick phenotype of the double Scgas1Δ bgl2Δ mutant suggested that β-(1,3)-glucan branching is essential. In addition to ScGas1p, GH72 family ScGas2p and Aspergillus fumigatus Gel4p, having CBM43 in their sequences, showed dual β-(1,3)-glucan elongating and branching activity. Our report identifies the fungal cell wall β-(1,3)-glucan branching mechanism. The essentiality of β-(1,3)-glucan branching suggests that enzymes involved in the glucan branching could be exploited as antifungal targets.
format	article
author	Vishukumar Aimanianda Catherine Simenel Cecile Garnaud Cecile Clavaud Rui Tada Lise Barbin Isabelle Mouyna Christoph Heddergott Laura Popolo Yoshikazu Ohya Muriel Delepierre Jean-Paul Latge
author_facet	Vishukumar Aimanianda Catherine Simenel Cecile Garnaud Cecile Clavaud Rui Tada Lise Barbin Isabelle Mouyna Christoph Heddergott Laura Popolo Yoshikazu Ohya Muriel Delepierre Jean-Paul Latge
author_sort	Vishukumar Aimanianda
title	The Dual Activity Responsible for the Elongation and Branching of β-(1,3)-Glucan in the Fungal Cell Wall
title_short	The Dual Activity Responsible for the Elongation and Branching of β-(1,3)-Glucan in the Fungal Cell Wall
title_full	The Dual Activity Responsible for the Elongation and Branching of β-(1,3)-Glucan in the Fungal Cell Wall
title_fullStr	The Dual Activity Responsible for the Elongation and Branching of β-(1,3)-Glucan in the Fungal Cell Wall
title_full_unstemmed	The Dual Activity Responsible for the Elongation and Branching of β-(1,3)-Glucan in the Fungal Cell Wall
title_sort	dual activity responsible for the elongation and branching of β-(1,3)-glucan in the fungal cell wall
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/7c42e07ba98241b8ae37693ee5b262ae
work_keys_str_mv	AT vishukumaraimanianda thedualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT catherinesimenel thedualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT cecilegarnaud thedualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT cecileclavaud thedualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT ruitada thedualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT lisebarbin thedualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT isabellemouyna thedualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT christophheddergott thedualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT laurapopolo thedualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT yoshikazuohya thedualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT murieldelepierre thedualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT jeanpaullatge thedualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT vishukumaraimanianda dualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT catherinesimenel dualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT cecilegarnaud dualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT cecileclavaud dualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT ruitada dualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT lisebarbin dualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT isabellemouyna dualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT christophheddergott dualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT laurapopolo dualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT yoshikazuohya dualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT murieldelepierre dualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall AT jeanpaullatge dualactivityresponsiblefortheelongationandbranchingofb13glucaninthefungalcellwall
_version_	1718427346665472000

The Dual Activity Responsible for the Elongation and Branching of β-(1,3)-Glucan in the Fungal Cell Wall

Ejemplares similares